Figure 2.

Growth of 4T1 cancer cells in vitro and in vivo upon GPD2 modulation. (A) Relative numbers of 4T1 and 4T1 ρ0 cells after treatment of KM04416 at different concentrations (0, 5, 10, and 20 µM) for 48 h. Both 4T1 and 4T1 ρ0 cell numbers were normalized by each paired control (0 µM KM04416). The cell numbers were counted with an automated cell counter. (B) Protein expression of GPD2 in 4T1 and 4T1 GPD2 KO cells as detected by Western blot analysis. (C) GPD2 enzyme activity in 4T1 and 4T1 GPD2 KO cells. (D) Growth rates of 4T1 and 4T1 GPD2 KO cells by counting cell numbers in 5 days of incubation. (E) Photographic image of clonogenic assay for 4T1 and 4T1 GPD2 KO cells. (F) Representative image of wound healing assay for 4T1 and 4T1 GPD2 KO cells (0 h and 48 h). (G) Tumor growth of syngeneic graft model of 4T1 and 4T1 GPD2 KO (1) cells. The size of the tumor was measured once every week with a caliper. (H) Photographic image of grafted tumors in (G). The tumors were extracted after 4 weeks, and photographed. (I) Weight of the extracted graft tumors in (H). Data were obtained from three biologically independent samples or four mice in each group unless indicated otherwise. The p-value was calculated by comparing the experimental group with 4T1 control group using a two-tailed unpaired Student's t-test. The “*” in the graphs indicates statistically significant difference (“*”: p < 0.05; “**”: p < 0.005; “***”: p < 0.0005).